农杆菌介导四季秋海棠遗传转化体系建立  

Establishment of Agrobacterium-mediated genetic transformation system for Begonia semperflorens

作　　者：李逸菲 王瑞博 夏惠婷 任璞玉 赵丹丹 秦肖洁 李永华[1] 张开明[1] LI Yifei;WANG Ruibo;XIA Huiting;REN Puyu;ZHAO Dandan;QIN Xiaojie;LI Yonghua;ZHANG Kaiming(School of Landscape Architecture and Art,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南农业大学风景园林与艺术学院,郑州450002

出　　处：《东北农业大学学报》2023年第5期19-27,共9页Journal of Northeast Agricultural University

基　　金：国家自然科学基金面上项目(32172622)。

摘　　要：以四季秋海棠‘超级奥林匹克’叶片为外植体,探索愈伤组织对卡那霉素筛选浓度;将外源基因表达载体通过农杆菌介导进行遗传转化,探索优化预培养、侵染液浓度(OD600)、侵染时间和共培养时间。结果表明,卡那霉素最适浓度为25 mg·L^(-1)。将四季秋海棠BsRbohD外源目的基因导入愈伤组织,经转基因植株分子鉴定,共获得两组不同遗传转化条件阳性植株共12株,分别为:预培养2 d、侵染液浓度OD_(600)=0.7、侵染时间10 min、共培养4 d,阳性率为11.1%;预培养1 d、侵染液浓度OD_(600)=0.7、侵染时间8 min、共培养4 d,阳性率为58.8%。研究初步建立农杆菌介导四季秋海棠稳定遗传转化体系,为深入开展分子设计育种与基因组学研究提供技术支持。Using the leaves of Begonia semperflorens'Super Olympia'as the explants,the screening concentration of kanamycin in callus was explored;Agrobacterium tumefaciens-mediated genetic transformation of foreign gene expression vectors was performed to investigate and optimize the pre-culture,infection concentration(OD600),infection time and co-culture time.The results showed that the optimal kanamycin concentration was determined to be 25 mg·L^(-1).The exogenous target gene BsRbohD was introduced into the calli of B.semperflorens.Following transgenic plant molecular identification,a total of 12 positive plants with two groups of different genetic transformation circumstances were obtained as the followings:after 2 d of pre-culture,10 min of infection solution concentration OD_(600)=0.7,and 4 d of co-culture,the positive rate was 11.1%.After 1 d of pre-culture,8 min of infection solution concentration OD_(600)=0.7,and 4 d of co-culture,the positive rate was 58.8%.The study had preliminarily established a stable genetic transformation system for B.semperflorens mediated by A.tumefaciens,providing technical supports for in-depth molecular design breeding and genomics researches.

关 键 词：四季秋海棠 组织培养 外植体 遗传转化体系 

分 类 号：S682.1[农业科学—观赏园艺] Q785[农业科学—园艺学]